• The Plant Pathology Journal
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• v.25 no.2
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• pp.184-188
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• 2009

Identification of differently expressed genes under specific tissues and/or environments provides insights into the nature and underlying mechanisms of cellular processes. Although cDNA-AFLP (Amplified Fragment Length Polymorphism) is a powerful method for analyzing differentially expressed genes, its use has been limited to the requirement of radioactive isotope use and the difficulty of isolating the bands of interest from a gel. Here, we describe a modified method for cDNA-AFLP that uses a fluorescence dye for detection and isolation of bands directly from a small size polyacrylamide gel. This method involves three steps: (i) preparation of cDNA templates, (ii) PCR amplification and differential display, and (iii) identification of differentially expressed genes. To demonstrate its utility and efficiency, differentially expressed genes during vegetative growth and appressorial development of Magnaporthe oryzae were analyzed. This method could be applied to compare gene expression profiles in a diverse array of organisms.

• Proceedings of the Microbiological Society of Korea Conference
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• 2002.10a
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• pp.15-21
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• 2002

Terminal-restriction fragment length polymorphism (T-RFLP) analysis has been optimized by using in vitro model community composed of genomic DNAs of known bacterial strains and has been applied to assess the bacterial community structure in marine sediments. The specific fluorescence-labeled terminal restriction fragments (T-RFs) between 39 and 839 base long specifying each strain were precisely measured for known bacterial strains. The addition of a co-solvent (dimethylsulfoxide or glycerol) into PCR reactions has reduced differential PCR amplification. Comparative bacterial community structure was investigated for pristine and polluted sediments. A complex T-RFLP pattern showing complex bacterial community structure was obtained in the pristine sediment, whereas simple T-RFLP pattern (low bacterial diversity) was shown in polluted sediments where caged aquaculture has been conducted for several years. The results of T-RFLP analysis were compared with that of cloning and sequencing 16S rDNA clones from the same sediments. Sequence analysis of 16S rDNA clones (72) of the pristine sediment revealed a diverse collection of lineages, largely of the class Proteobacteria ($6\%$ alpha subdivision, $46\%$ gamma subdivision, $13\%$ delta subdivision, and $3\%$ epsilon subdivision), Nitrospina $(8\%)$, high G+C gram positive $(8\%)$, Verrucomicrobia $(7\%)$, and Planctomycetes $(6\%)$. In the contaminated sediments, 17 $(59\%)$ of the 16S rDNA clones (29) were related to Campylobacter and symbiont of Rimicaris exoculata belonging to epsilon subdivision of Proteobacteria. The results obtained indicated that T-RFLP analysis is a rapid and precise technique for comparative bacterial community analysis.

• Mycobiology
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• v.37 no.3
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• pp.225-229
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• 2009

Single spore isolates of Plasmodiophora brassicae e4 and e9 obtained from diseased Chinese cabbage were identified as race 4 and race 9, respectively, by the Williams' differential variety set. To confirm the possibility of variation in same generation and progeny of a single spore isolate of P. brassicae, random amplified polymorphic DNA (RAPD) analysis was conducted using the URP 3, 6 and OPA 7 primers. There was no difference in band type at each part of the gall of Chinese cabbage obtained by inoculation of e4 and e9 and amplification using the URP 3 and 6 primers when the same generation was analyzed. In addition, the progeny analysis, which was expanded to the third generation and conducted using the URP 3 and OPA 7 primers, revealed no differences in the band type of the e4 isolate. Based on these results, the single spore isolate of P. brassicae was genetically stable.

• Korean Journal of Veterinary Research
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• v.38 no.1
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• pp.77-83
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• 1998

돼지 생식기호흡기증후군(porcine reproductive and respiratory syndrome, PRRS) 바이러스가 웅돈에 감염되었을 경우에는 정충의 기형 등 정액의 질 저하와 더불어 정액에 바이러스가 함유되어 있어 종부시 모돈에 바이러스를 전파하는 것으로 알려져 있다. 감염된 웅돈의 정액으로 인공수정을 실시할 경우 농장의 모돈 전체에 순식간에 바이러스를 전파하여 막대한 피해를 유발할 가능성이 높다. 따라서 감염웅돈을 신속히 검색하여 격리함으로써 피해를 사전에 방지해야 하며, 이를 위해서 웅돈의 감염여부를 신속히 확진하는 진단법의 개발이 필요한 실정이다. PRRS의 진단을 위해서는 바이러스학적인 진단법으로는 바이러스 분리동정, 혈청학적인 방법으로 바이러스 분리동정이 필수적이나 검사시간이 많이 소요되고, 분리동정 자체가 까다로운 단점이 있다. 본 연구에서는 웅돈의 정액내에서 PRRSV 바이러스에 대한 유전자를 RT-PCR법으로 증폭하는 방법을 개발하였으며, 진단의 민감도를 높이기 위하여 Nested PCR법으로 재확인 할 경우, 바이러스의 역가가 $1TCID_{50}$만 함유되어 있어도 진단이 가능한 조건을 확립하였다. 이 방법을 이용할 경우 웅돈의 정액시료에 대한 PRRS 바이러스 감염여부를 신속, 정확하게 검색하여 감염웅돈을 통한 PRRS바이러스의 전파를 미리 차단할 수 있으므로 PRRS방제에 효과적으로 이용될 것으로 사료된다.

• The Transactions of The Korean Institute of Electrical Engineers
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• v.65 no.5
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• pp.857-861
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• 2016

In this study, we proposed a new method that can be measure ECG (Electrocardiography) and PPG (Photoplethysmography) in realtime on the site of the wrist for check the state of health in daily life. For convenience measurement of ECG the lead I method was used on the wrist, and omit the reference junction ECG I was measured in the right hand and the left hand of the potential difference. Then the measured electrocardiogram was amplified by the differential amplifier and the signals were passed HPF, LPF, and BPF filters. For removing the PPG's noise from the Motion artifact and temperature, we apply the reflective photoelectric volume pulse wave measurement method using green LED as a light source. The circuits was designed to be able to check the waveform using higher active amplification method at weak signals. For the validation of our device, the measured signals were compared with E2-KIT on same time. The results shows that the error does not exceed the maximum one, most of the data is confirmed to be issued Peak inspection of the same number.

• Journal of the Korean Chemical Society
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• v.14 no.4
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• pp.327-332
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• 1970

A simple and convenient device for deflection-type direct reading the variations in electrolytic conductance is described and applied to the analysis of sulfur by combustion-$H_2O_2$ oxidation method. The apparatus consisted of a high resistance-ratio bridge in which the other adjacent arms are the differential cells. By adopting unusually high a-c voltage source for the bridge excitation, the a-c method for unbalanced bridge is established, decreased sensitivity owing to reduced bridge factor, 0.01, is overcome and also the absolute sensitivity and linearity are greatly improved. Over 50% variations in impedance of the balanced cell, within 1% deviation from the linearity can be attained with a volt (rms)order of output which was detected directly with VTVM without further amplification. Analysis of the bridge shows that these useful features are natural result of the constant current character of the high source impedance generator and the performance of the device agreed with the theoretical predictions. A standard procedure for the rapid analysis of sulfur using the bridge is also given, the analytical accuracy was approximately 1%. A determination takes not more than 5 minutes.

• Journal of Magnetics
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• v.14 no.1
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• pp.42-46
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• 2009

The measurement of external magnetic field orientation using Giant Magneto Impedance (GMI) sensors has been performed. A soft magnetic alloy of $Co_{30}Fe_{34}Ni_{36}$ was electroplated on a Si wafer with a CoFeNi seed layer. V-shaped microwire patterns were formed using a conventional photolithography process. An external magnetic field was generated by a rectangular AlNiCo permanent magnet. The reference direction was defined as the external magnetic field direction oriented in the middle of 2 GMI devices. As the orientation of the magnetic field deviated from the reference direction, variation in the field component along each device introduced voltage changes. It was found that, by taking the voltage difference between the left and right arms of the Vshaped device, the nonlinearity of each device could be significantly reduced. The fabricated angle sensor had a linear range of approximately $70^{\circ}$ and an overall sensitivity of approximately 10 mV.

• Advances in Computational Design
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• v.4 no.3
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• pp.307-326
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• 2019

A low computational cost semi-analytical method is developed, based on eigenfunction expansion, to study the vibration of rectangular plates subjected to a series of moving sprung masses, representing a bridge deck under multiple vehicle or train moving loads. The dynamic effects of the suspension system are taken into account by using flexible connections between the moving masses and the base structure. The accuracy of the proposed method in predicting the dynamic response of a rectangular plate subjected to a series of moving sprung masses is demonstrated compared to the conventional rigid moving mass models. It is shown that the proposed method can considerably improve the computational efficiency of the conventional methods by eliminating a large number of time-varying components in the coupled Ordinary Differential Equations (ODEs) matrices. The dynamic behaviour of the system is then investigated by performing a comprehensive parametric study on the Dynamic Amplification Factor (DAF) of the moving loads using different design parameters. The results indicate that ignoring the flexibility of the suspension system in both moving force and moving mass models may lead to substantially underestimated DAF predictions and therefore unsafe design solutions. This highlights the significance of taking into account the stiffness of the suspension system for accurate estimation of the plate maximum dynamic response in practical applications.

• ALGAE
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• v.34 no.1
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• pp.57-70
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• 2019

DNA metabarcoding is currently used for large-scale taxonomic identification to understand the community composition in various marine ecosystems. However, before being widely used in this emerging field, this experimental and analytic approach still has several technical challenges to overcome, such as polymerase chain reaction (PCR) bias, and lack of well-established metabarcoding markers, a task which is difficult but not impossible to achieve. In this study, we present an adapted PCR-free small-organelles enriched metagenomics (SoEM) method for marine biodiversity assessment. To avoid PCR bias and random artefacts, we extracted target DNA sequences without PCR amplification from marine environmental samples enriched with small organelles including mitochondria and plastids because their genome sequences provide a valuable source of molecular markers for phylogenetic analysis. To experimentally enrich small organelles, we performed subcellular fractionation using modified differential centrifugation for marine environmental DNA samples. To validate our SoEM method, two marine environmental samples from the coastal waters were tested the taxonomic capturing capacity against that of traditional DNA metabarcoding method. Results showed that, regardless of taxonomic levels, at least 3-fold greater numbers of taxa were identified in our SoEM method, compared to those identified by the conventional multi-locus DNA metabarcoding method. The SoEM method is thus effective and accurate for identifying taxonomic diversity and presents a useful alternative approach for evaluating biodiversity in the marine environment.

• Microbiology and Biotechnology Letters
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• v.47 no.3
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• pp.423-433
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• 2019

Midgut microbiota is known to play a fundamental role in the biology and physiology of the agricultural pest, Spodoptera litura. This study reports the difference in the larval midgut microbiota of field-caught and laboratory-reared populations of S. litura by performing 16S rDNA amplicon pyrosequencing. Field populations for the study were collected from castor crops, whereas laboratory-reared larvae were fed on a regular chickpea based diet. In total, 23 bacterial phylotypes were observed from both laboratory-reared and field-caught caterpillars. Fisher's exact test with Storey's FDR multiple test correction demonstrated that bacterial genus, Clostridium was significantly abundant (p < 0.05) in field-caught larvae of S. litura as compared to that in the laboratory-reared larvae. Similarly, bacterial genera, such as Bradyrhizobium, Burkholderia, and Fibrisoma were identified (p < 0.05) predominantly in the laboratory-reared population. The Bray-Curtis dissimilarity matrix depicted a value of 0.986, which exhibited the maximum deviation between the midgut microbiota of the laboratory-reared and field-caught populations. No significant yeast diversity was seen in the laboratory-reared caterpillars. However, two yeast strains, namely Candida rugosa and Cyberlindnera fabianii were identified by PCR amplification and molecular cloning of the internal transcribed space region in the field-caught caterpillars. These results emphasize the differential colonization of gut residents based on environmental factors and diet.